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B cells that produce anti–aquaporin-4 (AQP4) antibodies play a crucial role in neuromyelitis optica spectrum disorder (NMOSD) pathogenesis. We previously reported that naïve B (NB) cells from patients with NMOSD, unlike those from healthy controls, exhibit transcriptional changes suggesting the adoption of an antibody-secreting cell (ASCs) phenotype. CD25+ NB cells, whose numbers are increased in NMOSD patients, have a greater capacity to differentiate into ASCs than do CD25- NB cells. Here, we attempted to establish novel B cell subset cell lines from patients with NMOSD to enable molecular analysis of their abnormalities. We generated Epstein-Barr virus–immortalized lymphoblastoid cell lines (LCLs) from CD25+ NB, CD25- NB, and switched memory B (SMB) cells. All LCLs largely maintained the features of the original cell type in terms of cell surface marker expression and could differentiate into ASCs. Notably, CD25+ NB–LCLs derived from patients with NMOSD exhibited a greater capacity to differentiate into SMB-LCLs than did CD25- NB–LCLs derived from patients with NMOSD, suggesting that the established LCLs maintained the characteristics of cells isolated from patients. The LCLs established in this study are likely to be useful for elucidating the mechanism by which cells that produce anti-AQP4 antibodies develop in NMOSD.

Although Janus kinase inhibitor (JAKi) therapy is used for patients with autoimmune diseases (AD), one safety concern, interstitial lung disease (ILD), is life-threatening. We evaluated actual usage of JAKi and safety upon JAKi treatment, in an epidemiological retrospective cohort study utilizing the electronic medical record database in Japan. Among 391,565 AD patients, we analyzed data of new-users receiving JAKi or tumor necrosis factor alpha inhibitor (TNFi)/biologics during the period July 2013–May 2022. ILD (ICD10: J70.2, J70.3, J70.4 and J84) criteria were defined: new-ILD (1) and new-ILD (2) which differed in the latter’s prompter therapeutics cessation upon ILD development. We analyzed ILD occurrence and death, ILD cumulative incidence by the Kaplan–Meier method, and hazard ratio (HR) by the Cox model, for 957 JAKi and 3931 TNFi users. JAKi use has become widespread amidst additional drug-development. Among JAKi users, two-year new-ILD (2) incidence, at 1.4%, was higher than for TNFi users (risk ratio: new-ILD (2) 1.75, death 2.31). Cumulative incidence (2.9% in 20.48 days) was also significantly higher (log-rank test p = .013, HR 2.23 (95% CI 1.16–4.27)); risk factors estimated by HR included JAKi (2.14), rheumatoid arthritis (4.94), diabetes mellitus (2.67) and cerebrovascular disease (2.86). ILD screening is essential.

Myocarditis is an uncommon presentation in systemic sclerosis. We report the case of a patient who developed myocarditis on the initial diagnosis of systemic sclerosis and was successfully managed with optimal medical treatment, including immunosuppression. This case was able to raise awareness in recognizing the heart as an organ of involvement in systemic sclerosis and showed the importance of screening for heart involvement in the management of systemic sclerosis. The management of primary myocardial involvement in systemic sclerosis was also reviewed.

Nonsteroidal anti-inflammatory drugs (NSAIDs) are widely used for their analgesic, antipyretic, and anti-inflammatory properties. However, they are also associated with a broad spectrum of hypersensitivity reactions, ranging from mild cutaneous manifestations to severe systemic responses. The complex nature of these reactions and their underlying mechanisms pose challenges in diagnosis and management. We review the clinical evidence to six distinct clinical phenotypes of NSAID hypersensitivity, including: NSAID-induced urticaria/angioedema or anaphylaxis (SNIUAA), NSAID-exacerbated respiratory disease (NERD), NSAID-exacerbated cutaneous disease (NECD), NSAID-induced urticaria/angioedema (NIUA), food-dependent NSAID-induced hypersensitivity reactions (FDNIH), and selective NSAID-induced delayed reactions (SNIDR). This review aims to provide a comprehensive review of NSAID hypersensitivity reactions based on clinical phenotyping, which can aid in understanding and managing these adverse events.

Objective: To assess the predictive accuracy of the EULAR definition of clinically suspected arthralgia (CSA) for progression to rheumatoid arthritis (RA) in patients presenting with arthralgia without arthritis. Methods: This prospective study recruited 100 patients with arthralgia but no clinical arthritis. Patients were evaluated at baseline and followed up every 6 months for 2 years. The association of the presence of [Formula: see text] and [Formula: see text] out of seven CSA parameters at baseline with RA development was investigated. The association of individual CSA parameters with RA development was also evaluated. Results: Twenty-one of 100 patients (21%) developed RA. The proportion of patients meeting [Formula: see text] CSA criteria was higher in those who developed RA (66.7%) compared to those who did not (39.2%, [Formula: see text]). Similarly, a higher proportion of patients who developed RA met [Formula: see text] CSA criteria (47.6%) compared to those who did not (20.3%, [Formula: see text]). The sensitivity and specificity of [Formula: see text] CSA parameters were 71.4% and 69.6%, and for [Formula: see text] CSA parameters were 42.9% and 87.3%, respectively. “Symptom duration [Formula: see text] year” and “difficulty holding a fist” were significantly associated with RA development. A simplified criteria using these two items provided a similar area under receiver operating characteristic (ROC) curve (0.747 vs. 0.676) compared to the original criteria. Conclusion: Meeting EULAR CSA criteria was significantly associated with progression to RA in patients with arthralgia. Symptom duration [Formula: see text] year and difficulty holding a fist were the most predictive individual parameters. Continuous monitoring using the CSA criteria, and probably a simplified version, could help identify high-risk individuals for RA development.

Objective: To describe the clinical profile and predictors of mortality of antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) patients in Hong Kong. To compare the accuracy of the latest Five-Factor Score (FFS-2009) and the Birmingham Vasculitis Activity Score (BVAS) in prediction of survival with this local cohort. Methods: A retrospective observational study on newly diagnosed AAV patients, from January 1, 2011 to March 31, 2022, managed in the Kowloon West Cluster (KWC) hospitals in Hong Kong. Demographic and baseline characteristics, clinical profile, and treatment profile were reviewed. Factors associated with mortality were analyzed with the Cox proportional hazards model. The performances of FFS and BVAS in mortality prediction were analyzed by receiver operating characteristic (ROC) curves. Results: A total of 83 AAV patients were included in the study. The median age was 70.5 years at diagnosis. Microscopic polyangiitis (MPA; 69.9%) was the most common AAV subtype. The median FFS and BVAS were 2 and 20, respectively. The overall mortality was 45.6% across the study period. Multivariate Cox regression identified age at diagnosis (HR 1.043, [Formula: see text]), stabilized peak serum creatinine (HR 1.002, [Formula: see text]), hemoglobin level (HR 0.754, [Formula: see text]), cardiac involvement (HR 3.862, [Formula: see text]), and use of maintenance therapy (HR 0.261, [Formula: see text]) as independent predictors of overall survival. Both FFS and BVAS were significant predictors of overall survival. The areas under the curve (AUC) of ROC curves suggested FFS was a good prediction tool for early mortality in 1 year, with an AUC value of 0.874. Conclusion: Despite the advances in treatment, AAV still carried significant morbidities with high mortality. Clinical predictors and existing scoring systems showed good predictive power on mortality.

Idiopathic inflammatory myopathy (IIM) poses elevated risk of cardiovascular event and mortality, similar to other autoimmune rheumatic diseases. We conducted a cross-sectional study to examine the prevalence and risk factor of subclinical coronary artery disease (CAD) in patients with IIM, using CT coronary angiogram (CTCA). The prevalence of obstructive CAD and CAD in IIM (13.3% and 66.7%, respectively) were significantly higher than age and sex-matched controls (0% and 30%, respectively, both [Formula: see text]). Diabetes mellitus and calcium calcification score [Formula: see text] units were found to be the independent predictors of obstructive CAD. Screening of high-risk patients with aggressive treatment of cardiovascular risk factors should be considered in managing IIM patients.

A 38-year-old lady was diagnosed with systemic lupus erythematosus (SLE) and alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor (AMPAR) autoimmune encephalitis, fulfilling both classification criteria during her index admission. She was subsequently diagnosed with seropositive myasthenia gravis (MG) with a typical clinical presentation. Her SLE and AMPAR encephalitis were successfully treated with immunosuppressive therapy. Her MG responded well to anti-cholinesterase inhibitors. We report a distinctive case with the association of three rare autoimmune diseases in an individual.

Abstract Type 2 diabetes mellitus (T2DM) is a metabolic disorder characterized by hyperglycemia resulting from defects in insulin secretion and/or insulin action. Increasing evidence suggests that inflammation played an important role in the pathogenesis of T2DM. Prospective studies on the link between immunoglobulins concentrations and the risk of T2DM in adults are limited. We developed a cohort study including 7,093 adults without T2DM history. The incidence of T2DM was 16.45 per 1,000 person-years. Compared with the lowest quartiles, the hazard ratios (95% confidence intervals) of T2DM for the highest quartiles of IgG, IgE, IgM and IgA were 0.64 (0.48–0.85), 0.94 (0.72–1.23), 0.68 (0.50–0.92) and 1.62 (1.24–2.11) (P for trend was

Abstract Purpose Primary Sjogren's syndrome (pSS) is a prevalent autoimmune disease. The immune dysregulation it causes often leads to the development of diffuse large B-cell lymphoma (DLBCL) in clinical practice. However, how it contributes to these two disorders at the molecular level is not yet known. This study explored the potential molecular mechanisms associated with the differences between DLBCL and pSS. Patients and methods Gene expression matrices from discovery cohort 1, discovery cohort 2, and the validation cohort were downloaded from the GEO and TCGA databases. Weighted gene coexpression network analysis (WGCNA) was performed to identify the coexpression modules of DLBCL and pSS in discovery cohort 1 and obtain shared genes. GO and KEGG enrichment analyses and PPI network analysis were performed on the shared genes. Immune-related genes (IRGs) were intersected with shared genes to obtain common genes. Afterward, common genes were identified via machine learning methods. The immune infiltration analysis, miRNA-TF-hub gene regulatory chart, gene interactions of the hub genes, and gene‒drug target analysis were performed. Finally, STAT1 was identified as a possible essential gene by the above analysis, and immune infiltration and GSEA pathway analyses were performed in the high- and low-expression groups in discovery cohort 2. The diagnostic efficacy of the hub genes was assessed in the validation cohort, and clinical samples were collected for validation. Results By WGCNA, one modular gene in each group was considered highly associated with the disease, and we obtained 28 shared genes. Enrichment analysis revealed shared genes involved in the viral response and regulation. We obtained four hub genes (ISG20, STAT1, TLR7, and RSAD2) via the algorithm. Hub genes and similar genes are primarily involved in regulating type I IFNs. The construction of a miRNA-TF-hub gene regulatory chart revealed that hsa-mir-155-5p, hsa-mir-146b-5p, hsa-mir-21-3p, and hsa-mir-126-3p play essential roles in both diseases. Hub genes were differentially expressed in B-cell memory according to immune infiltration analysis. Hub genes had a strong diagnostic effect on both diseases. STAT1 plays an essential role in immune cells in both diseases. Conclusion We identified hub susceptibility genes for DLBCL and pSS and identified hub genes and potential therapeutic targets that may act as biomarkers.

Abstract Objective Fibromyalgia (FM) subjects are treated with antidepressant agents; in most cases, these drugs lose efficacy or have adverse effects. Ketamine is an anesthetic drug used in FM in some studies. This article aims to systematically review the safety and efficacy of ketamine in fibromyalgia (FM) patients. Materials and methods We systematically searched articles on FM and ketamine published at Pubmed from 1966 to 2021. This study was registered at PROSPERO. Results There were only 6 articles published in this field, with a total of 115 patients. The female sex was predominant (88 to 100%). The age varied from 23 to 53 years old. Disease duration ranged from 1 month to 28 years. The dosage of ketamine changed from 0.1 mg/kg-0.3-0.5 mg/kg in intravenous infusion (4/5) and subcutaneous application (1/5). Regarding outcomes, the Visual analog scale (VAS) before ketamine was from 59 to 100 mm and after treatment from 2 to 95 mm. Most short-term studies had a good response. Only the study with 8 weeks of follow-up did not observe a good response. Side effects were common; all appeared during the infusion and disappeared after a few minutes of the ketamine injection. Conclusions The present study demonstrates the effectiveness and safety of ketamine in FM patients in the short term. Although, more studies, including long-term follow-up studies, are still needed.

Abstract Background Age-dependent immune responses to coronavirus disease 2019 (COVID-19) vaccinations and breakthrough infections (BIs) in young and middle-aged individuals are unclear. Methods This nationwide multicenter prospective cohort study analyzed immune responses in participants of the ChAdOx1 (ChAd)-ChAd-mRNA vaccine group using cytometry by time-of-flight, anti-spike protein antibody (Sab) and anti-nucleocapsid antibody (Nab) titers, plaque reduction neutralization tests (PRNTs), and interferon-gamma (IFN-γ) release assays at various time points. Results We evaluated 347 participants with an average age of 38.9 ± 9.4 years (range: 21–63). There was a significant inverse correlation between age and Sab levels after the second dose (slope − 14.96, P = 0.032), and this was more pronounced after the third dose (slope − 208.9, P

Abstract Background Effective management of antiretroviral therapy (ART) is crucial in combating the global HIV pandemic. This study, the first of its kind in Yemen, investigates the rate and determinants of switching from first-line to second-line ART among people living with HIV (PLWH) in Aden City, Yemen. Methods A retrospective cohort study was conducted using data from PLWH who started first-line ART at Al-Wahda Hospital from 2007 to May 2022. PLWH in prevention of mother-to-child transmission (PMTCT) programs, those already on second-line ART at enrollment, and those with less than 3 months of follow-up were excluded. Cumulative incidence curves and multivariable proportional hazards models were used to identify factors associated with switching, considering death and loss to follow-up as competing risks. Analyses were carried out using IBM SPSS version 26. Results Out of 149 PLWH, 18 (12.1%) switched to second-line ART with a cumulative incidence rate of 1.8 per 100 person-years. Significant factors for switching included being older than 33 years (HR: 1.45, 95% CI: 1.12–1.89), having WHO stage 3 disease (HR: 1.58, 95% CI: 1.21–2.06), and being on a TDF-FTC-EFV-based first-line regimen (HR: 1.35, 95% CI: 1.03–1.77). This switching rate is consistent with rates observed in other resource-limited settings, indicating it is neither exceptionally high nor low compared to similar contexts​. Conclusions The study highlights key factors associated with switching to second-line ART in Yemen, emphasizing the need for targeted interventions and continuous monitoring to enhance treatment outcomes. These findings are consistent with regional data from other resource-limited settings.

Abstract In 2022, a genotype IV (GIV) strain of Japanese encephalitis virus (JEV) caused an unprecedented and widespread outbreak of disease in pigs and humans in Australia. As no veterinary vaccines against JEV are approved in Australia and all current approved human and veterinary vaccines are derived from genotype (G) III JEV strains, we used the recently described insect-specific Binjari virus (BinJV) chimeric flavivirus vaccine technology to produce a JEV GIV vaccine candidate. Herein we describe the production of a chimeric virus displaying the structural prM and E proteins of a JEV GIV isolate obtained from a stillborn piglet (JEVNSW/22) in the genomic backbone of BinJV (BinJ/JEVNSW/22-prME). BinJ/JEVNSW/22-prME was shown to be antigenically indistinguishable from the JEVNSW/22 parental virus by KD analysis and a panel of JEV-reactive monoclonal antibodies in ELISA. BinJ/JEVNSW/22-prME replicated efficiently in C6/36 cells, reaching titres of >107 infectious units/mL - an important attribute for vaccine manufacture. As expected, BinJ/JEVNSW/22-prME failed to replicate in a variety of vertebrate cells lines. When used to immunise mice, the vaccine induced a potent virus neutralising response against JEVNSW/22 and to GII and GIII JEV strains. The BinJ/JEVNSW/22-prME vaccine provided complete protection against lethal challenge with JEVNSW/22, whilst also providing partial protection against viraemia and disease for the related Murray Valley encephalitis virus. Our results demonstrate that BinJ/JEVNSW/22-prME is a promising vaccine candidate against JEV.

Despite significant progress in basic and preclinical research into acute myeloid leukaemia (AML), the five-year survival rate for patients with AML remains poor, highlighting the urgent need for new combination therapies. Over the past decade, increased attention has been focused on identifying suitable immunotherapeutic strategies to combat AML, in particular targeting leukaemia cells and their precursors with cytokines. Targeted therapy is also an established approach for the treatment of AML. However, with the increasing number of treatment options, there are challenges in understanding how to select the most effective therapy and how to combine different drugs. Venetoclax is a targeted agent, a potent and highly selective inhibitor of B cell lymphoma protein-2 (BCL-2), one of the cell’s major anti-apoptotic proteins. Research into approaches to improve the treatment of AML remains challenging due to the limitations of experimental models. Despite improvements in ex vivo culture protocols, in vivo models remain the only way to study the inherently heterogeneous nature of AML and the influence of the microenvironment on leukaemia development. In our study, we show that in a xenograft mouse model of acute myeloid leukaemia in mice of the NSG-SGM3 line, there is an increase in serum IL-4 in response to venetoclax therapy. IL-4 has previously been shown to induce apoptosis in AML cells. These data provide new perspectives for the use of strategies based on the synergism of venetoclax and IL-4 in inducing apoptosis. The data also show an increase in serum human MCP-1 levels upon engraftment of OCI-AML-2 leukaemia cells in the serum of xenografted mice, which decreases after venetoclax therapy and may serve as a prognostic marker for the success of ongoing therapy. An obvious advantage of xenograft models in mice was the ability to separate the expression and secretion of murine cytokines and chemokines that determine the microenvironmental response from the cytokine profile of the human tumor cells themselves. Overall, our data suggest additional functional features of venetoclax action on tumor cells through the regulation of cytokine secretion and the prospect of using the immunodeficient mouse line NSG-SGM3 to test new approaches to the treatment of AML.

An unfavorable prognosis for ovarian cancer is associated with metastasis to the peritoneum and the formation of malignant ascites, which contains factors affecting the growth and survival of tumor cells. Molecular and functional analysis of ascites provides information both for clinical diagnosis and for understanding the mechanisms of progression and resistance in ovarian cancer. The aim of the study was to evaluate the levels of IL-8 and WISP1 in the acellular portion of ascites in advanced ovarian cancer. In 30 patients diagnosed with ascitic ovarian cancer stage III-IV according to FIGO, before treatment, the levels of IL-8 (Kit A-8762, Interleukin-8-ELISA-BEST, JSC Vector-Best, Russia) and WISP1 (Kit SEG895Hu CloudClone Corp., China) were determined (pg/mL). Based on the results of the effectiveness of chemotherapy according to the TP scheme, all patients were divided into the following groups: without relapse, relapse-free period up to 6 months – early relapse and progression during chemotherapy. Statistical processing was carried out using Statistica 13. Analysis of patient progression-free time was carried out using the Cox regression method, and the patient survival function was assessed using the Kaplan-Meier method (Jamovi 2.4.14). We found that the level of IL-8 in the acellular part of malignant ascites in ovarian cancer in patients without relapse is significantly lower than in the early relapse group (176.58 (139.68-217.01) pg/mL versus 320.43 pg/mL (250.49-369.81), p = 0.019). The level of WISP1 was significantly increased in the acellular part of ascites only in patients with progression during chemotherapy (980.51 (796.61-1524.15) pg/mL versus 770.55 (500.60- 1254.90) pg/mL in patients without relapse and 764.09 (581.55-823.38) pg/mL in patients with relapse). We found a positive strong Pearson correlation between IL-8 and WISP1 in ascites in a group of patients without relapse (r = 0.783, p = 0.012). In the multivariate version of Cox regression, the risk of relapse increases by 1.01 (1.01-1.02, p = 0.001) times with an increase in the level of IL-8 in the acellular part of ascites. When IL-8 levels in the acellular portion of ascites are above 225 pg/mL, the median progression-free time in patients with advanced ovarian cancer is 11.7 (5.2-18.2, 95% CI) months.Thus, increased levels of IL-8 and WISP1 in malignant ascites in ovarian cancer are associated with a shorter progression-free time. IL-8 in malignant ascites activates Wnt/β-catenin signaling in advanced ovarian cancer.

More and more new data, concerning extraoral bitter taste receptors (TAS2R), appear now. Current data on polymorphisms, expression patterns and form of TAS2R subtype 38 (TAS2R38), its molecular variants that differ in the degrees of sensitivity to ligands and their role in the pathogenesis of respiratory disorders are discussed in this review. The mechanism of signal transduction from taste receptors mediated by G-protein is shown. Participation of TAS2R38 in the local protective mechanisms in a ciliated epithelium of the respiratory tract and its activation by “quorum sensing” system molecules and its connection with the components of mucociliary clearance are presented. It has been shown that the action of the ligand on the TAS2R38 leads to the activation of NO synthase, followed by the production of nitric oxide (NO), which triggers a number of intracellular reactions leading to an increase in the rate of beating of the cilia of the ciliary epithelium, as well as having a direct antibacterial effect. TAS2R38 are also found on leukocytes, and its expression decreases with age, which can be considered as a component of the general aging of immunocompetent cells in the body. It is known that activation of TAS2R38 also enhances the phagocytic activity of macrophages, which is also mediated by the action of G-protein and cGMP. TAS2R receptors are also considered to be associated with allergic diseases, in particular – with bronchial asthma. A number of studies in groups of children with bronchial asthma revealed that the expression of most TAS2Rs was higher in children with severe bronchial asthma. Other studies have shown that patients with the eosinophilic variant of chronic rhinosinusitis have a higher levels of TAS2R38 expression in the upper respiratory tract compared to those with chronic rhinosinusitis without eosinophilia. To date, the functional significance of extraoral bitter taste receptors has not been fully studied. In the future, a large amount of research work remains to be done to finally understand the role of TAS2R in the pathogenesis of respiratory diseases.

Known methods of treating chronic polypous rhinosinusitis do not have a significant effect on the progressive nature of the disease. Recently, anticytokine therapy has been developed and introduced. However, the high cost of drugs and the complexity of their production make it necessary to search for other drugs with similar mechanisms of action. Such immunotropic drugs can be recombinant IFNα-2b and the γ-D-glutamyl-L-tryptophan. The aim of the study is to study the mechanism of action and evaluate the effectiveness of local application of IFNα-2b and γ-D-glutamyl-L-tryptophan in chronic polypous rhinosinusitis with concomitant bronchial asthma. Patients of the first group (31 people) were injected with IFNα-2b at a dose of 1 million units into the polypous tissue for five days. Patients of the second group (31 people) were injected with IFNα-2b at a dose of 1 million units and γ-D-glutamyl-L-tryptophan at a dose of 0.1 mg into the polypous tissue for five days. Patients were observed for a year. Before treatment and 1 month after treatment, a biopsy of polypous tissue was performed with analysis of morphological changes. Local administration of IFNα-2b and its combination with the γ-D-glutamyl-L-tryptophan into polypous tissue causes a decrease in the activity of T2 inflammation: decreased intercellular edema, tissue compaction, decreased the number of lymphocytes, eosinophils, plasma cells infiltrating the mucous membrane of polypous tissue. An analysis of the study results based on an assessment of the size of polyps showed that a month after therapy, the size of the polyps decreased, and when using IFNα-2b with γ-D-glutamyl-L-tryptophan, the size of the polyps decreased by approximately 70%, and the use only recombinant IFNα-2b by 40%. After 6 months, there was an increase in polyp tissue by 10-20% compared to the values after 1 month. After a year, the growth of polyps remained at the level of the values obtained after 6 months. The use of both IFNα-2b, and its combination with the γ-D-glutamyl-L-tryptophan, is a pathogenetically substantiated method of treating chronic polyposis rhinosinusitis, which allows its use as a conservative therapy for this disease.

Lymphocytes are key cells in inflammation. The realization of inflammation accompanied by the development of oxidative stress depends on metabolic processes occurring in blood lymphocytes. Experimental studies of molecular control of the redox status and apoptotic death of blood lymphocytes are relevant to study the role of lymphocytes in the pathogenesis of inflammation. The glutathione system plays a leading role in maintaining the redox status and oxidative modification of blood lymphocyte proteins. The study of molecular mechanisms of oxidative modification of proteins under the conditions of blocking glutathione synthesis is the basis for the targeting control of lymphocyte apoptosis. Inhibitory analysis is a molecular approach in experimental science used to study cellular metabolism by targeting specific stages of biochemical processes. The aim of the research was to determine the role of oxidative protein modification in redox regulation and cell death of blood lymphocytes when glutathione synthesis is inhibited during oxidative stress. The effect of exposure to the de novo glutathione synthesis inhibitor buthionine sulfoximine at a final concentration of 1 mM on the state of the glutathione system was studied in the experiment: content of reduced and oxidized glutathione, activity of glutathione reductase and glutathione peroxidase; on oxidative stress parameters: concentration of hydroxyl radical, reactive oxygen species, free SH-groups of proteins; on reversible and irreversible oxidative modification of proteins: content of glutathione bound to proteins, carbonyl derivatives of proteins, oxidized tryptophan and bityrosine; on realization and regulation of apoptotic death type: the number of annexin V+ cells and caspase-3 activity in blood lymphocytes. Blocking of de novo glutathione synthesis in blood lymphocytes was accompanied by the formation of oxidative stress, imbalance of glutathione system, changes in oxidative modification of proteins associated with the activation of apoptosis realization and completion. The obtained results indicate the participation of glutathione system components in reversible and irreversible oxidative modification of proteins, redox regulation and realization of apoptosis of blood lymphocytes. Therefore, modifying redox homeostasis through glutathionylation and carbonylation of cell proteins is a personalized apoptosis control mechanism.

Postischemic neuroinflammation is a critical pathophysiological process within the entire pattern of cerebral ischemia. It is characterized by microglial and astroglial activation and is accompanied by disturbances in the innate and adaptive immune response. The early damage of the blood-brain barrier (BBB) integrity is accompanied by the brain autoantigens release into circulation, in particular, the neurospecific protein S100B. According to recent experimental data, activated autophagy is associated with postischemic neuroinflammation, involved in its regulation and influences the outcome of the ischemic stroke (IS) acute period. Experimental evidence is provided for the autophagy involvement in the regulation of proinflammatory cytokines and chemokines production. The influence of activated autophagy on the pro- and anti-inflammatory cytokines balance in acute IS has been demonstrated. Purpose of the study: to quantitatively evaluate key autophagy biomarkers, the early biomarker of BBB damage S100B, pro- and anti-inflammatory cytokines in the dynamics of the IS acute period. To identify the relationship between autophagy and inflammation biomarkers, 112 patients with acute IS and 56 healthy persons were examined. Patients underwent dynamic clinical neurological examination and blood testing on the 1st, 7th and 14th days from the disease’s onset. The level of autophagy in peripheral blood leukocytes was determined by flow cytometry by assessing the intracellular expression of autophagy proteins LC3, p62 and mean fluorescence intensity of the Cyto-ID dye, which specifically recognizes active autophagosomes. Serum concentrations of TNFα, IL-1β, IL-4, IL-6, IL-8, IL-10, IL-18, neuropeptide S100B and autophagy biomarkers Beclin-1, LC3, p62 were determined by ELISA. A statistically significant increase in the studied biomarkers was found compared to the control group. The maximum increase in inflammation indicators and neuropeptide S100B was observed on the 1st, and autophagy biomarkers – on the 7th day of the disease. Established correlations indicate the participation of activated autophagy in the postischemic neuroinflammation regulation and its involvement in ischemic brain damage in the early stages of the IS acute period (days 1-7).

B cells play a crucial role in the pathogenesis of various diseases, such as autoimmune disorders, cancers, and infections. Unlike regulatory T cells, the anti-inflammatory capabilities of B cells have only recently garnered attention. Cytokines IL-10 and TGF-β are among the key secreted immunosuppressive factors, therefore studying the characteristics of their transcriptional regulation in B cells appears to be a relevant task. This study focuses on characterizing the promoter regions of IL10 and TGFB1 genes in immortalized B cell lines representing different developmental stages – Reh and Raji. To achieve this, we identified potential promoter regions guided by the epigenetic features of functional regulatory regions determined by bioinformatics methods of ChIP-Seq data analysis of chromatin marks in CD19+ lymphocytes. We examined the activity of selected promoters using reporter analysis in B cells. Additionally, we studied the impact of a single nucleotide polymorphism rs1800469 in the TGFB1 promoter, which is associated with the development of colorectal cancer, chronic obstructive pulmonary disease, and the risk of radiation fibrosis. Our results showed increased promoter activity of IL10 and TGFB1 in the Reh pro-B cells compared to the Raji mature B cells upon stimulation. Interestingly, the presence of the minor allele of rs1800469 led to enhanced TGFB1 promoter activity in the Reh cells. Higher activity of IL10 and TGFB1 promoters in acute lymphoblastic leukemia Reh cells may be associated with the increased immunosuppression, which is characteristic of this pathology. It is also possible that activation of pro-B cells Reh induces their differentiation into monocyte-like cells, which can be polarized into alternatively activated (M2) macrophages by autocrine TGF-β and IL-10. M2 macrophages can function as tumor-associated macrophages and contribute to the development of colorectal cancer. Moreover, increased levels of TGF-β in tissues increase the risks of fibrosis and decrease inflammation levels in chronic obstructive pulmonary disease.

Sarcoidosis is a multisystem inflammatory disease of unknown etiology characterized by the formation of non-caseating granulomas, most commonly in the lung tissue. It presents with two main forms: acute and chronic. Patients with chronic sarcoidosis tend to have a less favorable prognosis with a risk of developing lung fibrosis. Sarcoidosis development involves the activation of T cells, which release various chemokines and cytokines that stimulate the inflammatory process. The aim of our study was to investigate the role of the ratio between Th17 and Treg cells in the chronic course of sarcoidosis. We studied peripheral blood plasma samples from patients with chronic sarcoidosis (CS) (n = 101) and healthy individuals (HC) (n = 40). The diagnosis in CS patients was confirmed by histological methods. We determined the levels of Th17 and Treg (% of total lymphocytes) by flow cytometry. The concentration of cytokines (pg/ml) IL-17A and IL-10 was measured by multiplex analysis using Luminex xMap. Correlations between the Th17/Treg ratio and clinical parameters, including serum angiotensin-converting enzyme (sACE) activity level in the peripheral blood, forced expiratory volume in the first second (FEV1, %), fibrosis manifestations, and extrapulmonary manifestations were analyzed in CS patients. Our analysis revealed elevated levels of Th17 cells (p = 0.028) and decreased Treg levels (p = 0.026) in CS patients compared to healthy controls. This resulted in a significantly increased Th17/Treg ratio (p = 0.003) and IL-17A/IL-10 ratio (p < 0.001) in sarcoidosis patients. Furthermore, the Th17/Treg ratio positively correlated with sACE levels (p = 0.018), fibrosis manifestations (p = 0.019), and extrapulmonary manifestations (p = 0.016), and negatively correlated with FEV1% (p = 0.021). Our results indicate an increase in the Th17/Treg ratio, as well as the ratio of their main cytokines in patients with chronic sarcoidosis, which may emphasize their potential role as a diagnostic and prognostic biomarker of disease severity. At the molecular level, the balance between Treg and Th17 cells is maintained by the transcription factors Foxp3 and RORγt, which regulate the differentiation and function of these cells. Disruption of this balance in patients with chronic sarcoidosis may indicate a possible mechanism for disease progression.

The naked mole-rat (NMR) is a unique long-lived rodent with low cancer incidence. Understanding the molecular mechanisms that NMR evolved to control aging and tumorigenesis is important for biomedicine. It is commonly accepted that the immune system has essential functions in the tumor growth control in animals. In-depth study of the NMR immune system has recently begun, thus peculiarities of antitumor response in these animals remain undiscovered. However, it was shown that myelopoiesis predominates in NMR, therefore it can be assumed that the innate immune cells in the naked mole rat contribute to the successful control of the cancer incidence and tumor growth. This brief provides an overview of ongoing research into the properties of naked mole rat macrophages. Recent study shown that naked mole rat peritoneal macrophages are capable of acquiring an inflammatory phenotype (M1) but polarization into an anti-inflammatory phenotype (M2) under standard stimulus is limited. A more in-depth study using transcriptome sequencing and immunometabolic profiling in novel in vitro model of naked mole rat bone marrow macrophages suggested by our group revealed non-canonical features of M1 as well as M2 phenotypes of naked mole rat macrophages, which can be associated with the evolutionary adaptation of the species. Continued study of the different polarization conditions of naked mole rat macrophages is important to determine unique adaptations in NMR antitumor immunity.

The development of subclinical atherosclerosis in patients with rheumatoid arthritis (RA) is associated with chronic inflammation, one of the key mechanisms of which may be abnormal activation of macrophages.Objective: To assess the characteristics of pro-inflammatory activation of circulating monocytes in patients with early RA depending on the presence of subclinical atherosclerosis of the carotid arteries.The study included 60 patients (42 women and 18 men) with early RA without signs of cardiovascular disease. Atherosclerotic vascular disease was diagnosed by identifying carotid atherosclerotic plaques. Basal and stimulated monocyte lipolysaccharide (LPS) secretion was studied in initial monocyte cultures obtained by immunomagnetic separation from blood. Quantification of the cytokines TNFα and IL-1β was obtained in the culture fluid by ELISA. Proinflammatory activation of monocytes was calculated as the ratio of LPSstimulated and basal secretion.Atherosclerotic plaques of the carotid arteries were found in a third of RA patients; they were detected more often in men (50%) than in women (26%, p < 0.05). The carotid thickness of the intima media complex correlated with the level of total cholesterol (R = 0.20; p = 0.001) and ESR (R = 0.31; p = 0.03). In RA patients and subclinical carotid atherosclerosis, cultured blood monocytes demonstrated higher basal TNFα secretion (294.6 (185.3-778.2) vs 146.1 (27.9-79.9) pg/mL, p < 0.01) and low activation of TNFα (9.5±2.1 vs 19.8±3.9, p < 0.001) and IL-1β (6.1±2.3 vs 9.5±1.8, p = 0.03) compared with patients without lesions of the carotid arteries. In RA patients with carotid atherosclerotic plaques, a relationship was found between LPS-stimulated IL-1β secretion and the level of total blood cholesterol (R = 0.36, p = 0.01).Data were obtained on a more powerful inflammatory potential of peripheral blood monocytes in patients with early rheumatoid arthritis in the case of detection of the subclinical carotid atherosclerosis.

Activation of Toll-like receptors (TLRs) is one of the earliest indicators of functional activation of the innate immune system. Therefore, the development of drugs that stimulate the transcription of TLR/RLR genes and at the same time are “multi-target” drugs is an important task of modern immunopharmacology. In this regard, antiviral drugs that combine the properties of interferonogens and immunomodulators, which also include Cycloferon® and its analogues, are of great interest. The purpose of this study was to assess the expression of genes that determine the TLR/RLR signalling reactions of the innate immune system under the influence of immunomodulatory antiviral drugs based on acridoneacetic acid (Cycloferon® and Cycloferon L). The study was conducted using a model of immunocompetent cells: THP-1, differentiated by phorbol ester into macrophage-like cells. Gene expression analysis was performed using real-time polymerase chain reaction. The expression level of genes encoding TLR2, TLR3, TLR4, TLR7, TLR8, TLR9 and RIG-I was studied under the influence of the drugs Cycloferon® and Cycloferon L in three concentrations (156 μg/mL, 312 μg/mL and 625 μg/mL) on 1 hour, 4 hours and 24 hours. It was shown that the drug Cycloferon® at concentrations of 156, 312 and 625 μg/mL at 24 hours of exposure dose-dependently stimulated the expression of TLR2, TLR3, TLR4, TLR7, TLR8 receptor genes. A stable stimulation of the expression of RIG1 receptor genes was found upon exposure 4 hours to the drug in all studied concentrations. For the first time, it was revealed that the drug Cycloferon L stimulated a stable increase in the expression of TLR2, TLR3, TLR4, TLR7, TLR8 genes at an exposure period of 24 hours. Hereby, it was shown that the drugs Cycloferon® and Cycloferon L stimulated the expression of the TLR2, TLR3, TLR4, TLR7, TLR8 genes (and RIG1 for the drug Cycloferon), which are responsible for the synthesis of innate immune receptors.

Asthma is one of the most common chronic diseases in all age groups. Asthma has heterogeneous phenotypes with different etiologies. Many parameters are used to classify asthma, for example, the severity and level of flow control. The asthma phenotype is dependent on the state of the immune system, and innate immunity plays an important role in the susceptibility and pathophysiology of asthma. The complement system (CS) consists of a complex of protective proteolytic enzymes (including lectins). Ficolin-2 (L-ficolin) is one of the main opsonizing molecules of respiratory secretions and a protein of the lectin pathway of CS activation. Polymorphisms in the L-ficolin gene affect the level of expression which may be associated with a higher susceptibility to infections and viruses, as well as a predisposition to asthma.Aim: To study the distribution of polymorphisms rs17549193 and rs7851696 of the L-ficolin (FCN2) gene in children with asthma of varying severity.Russian children from the Children’s Allergy Center (Krasnoyarsk, Russia), aged from 8 to 18 years, were studied. Children with asthma were divided into groups depending on the severity of the disease in accordance with GINA-2023: mild (n = 146) and severe (n = 254). The comparison group included children of comparable age and gender without asthma, allergies or infections. DNA extraction from blood was performed using the sorbent method. Genotyping of polymorphisms rs17549193 and rs7851696 FCN2 was performed by real-time polymerase chain reaction.The results obtained provide distribution of the polymorphic variants FCN2 gene in the population of healthy Russian children and in children with a socially and economically important disease, namely asthma. The distribution of rs17549193 and rs7851696 FCN2 corresponds to the global Caucasoid populations. There were no statistically significant differences between asthma patients with varying degrees of severity of the disease and healthy ones in the studied sample.The results indicate an expansion of the sample and range of studied polymorphic genes of proteins of the lectin pathway of CS activation due to their importance for the prevention of severe forms of diseases, as well as their significance in the functioning of the immune system.

Cytokines and chemokines play dual – pro- and antioncogenic – roles in tumor progression. Targeted medications of monoclonal antibodies, anti-VEGF (bevacizumab) and anti-EGFR (cetuximab, panitumumab), are widely used in treatment of metastatic colorectal cancer (mCRC) and are prescribed in dependence upon presence or absence of mutations in the RAS gene. The aim of the study was to assess mCRC heterogeneity in the dependence upon presence or absence of mutation in RAS gene according to serum cytokine composition and its dynamics in the response to antitumor therapy using targeted medications of monoclonal antibodies. Levels of 20 cytokines were estimated by Multiplex analysis in serum of 50 patients with mCRC (25 KRAS+ and 25 KRAS- , who received anti-VEGF therapy, bevacizumab and anti EGFR therapy, cetuximab/ panitumumab respectively) before and after 4 courses of treatment. The results were analyzed separately in patients with complete, partial response and progression of the disease. The results showed that before the treatment in KRAS+ patients the levels of GM-CSF, IL-2, IL-5, IL-6, IL-7, IL-10, and IL-13 exceeded the ones in KRAS- patients; on the contrary, they had lower amounts of IL-8, IP-10, MIG, and MIP-1α. In patients who received anti-EGFR therapy and developed complete response, the increase of IL-15 and MIG along with a 2 to 3-fold decrease in GM-CSF, IL-2, IL-4, IL-6, IL-8, IL-17А, and МСР-1 was noted. Progression of the disease was observed in patients with initially low levels of the vast majority of the studied cytokines with dramatically elevation after non-effective anti-EGFR treatment. In patients having received anti-VEGF therapy, progression was followed by decrease in all of the studied cytokine and chemokine levels, while complete response resulted in decreases in IL-6, IL-5 and IL-10 (the last ones up to 0) and the increase of MIG. Thus, serum levels of cytokines in patients with mCRC were shown to be different in dependence of KRAS mutation; different response to targeted monoclonal antibodies may be reflected by the dynamics of serum cytokines` composition. Prevailing of many prooncogenic and proangiogenic cytokines in KRAS+ mCRC patients may be considered in terms of their unfavorable prognosis.

Patients with 22q11.2 deletion syndrome (DiGeorge syndrome) are characterized by a combination of a wide range of pediatric problems with an immunodeficiency. Defects are characterized by T cell lymphopenia, changes in the functions and subpopulation composition of T and B lymphocytes. Disturbances in lymphocyte homeostasis can lead not only to severe infectious diseases, but also to autoimmune complications, especially in older children. The purpose of this study was to compare the subpopulations of T and B lymphocytes with and without autoimmune complications and to search for prognostic signs that precede the development of complications. The study included 20 patients aged 10 to 18 years with a confirmed diagnosis of DiGeorge syndrome. The patients were divided into 2 groups, according to the presence or absence of autoimmune complications. Subpopulations of lymphocytes were assessed by flow cytometry. No statistically significant differences were found between CD3 T lymphocytes, CD4 T helper, CD8 T cytotoxic and subpopulations of T helper (p > 0.05). However, in the group of patients with autoimmune complications, a statistically significant decrease in CD45RA+ naïve T helper cells was detected, both in relative (p = 0.020) and absolute number (p = 0.025) and regulatory T cells (respectively, p = 0.020 and p = 0.007). Among B-lymphocyte in patients with autoimmune complications, a decrease in memory B cells in relative (p = 0.031) and absolute number (p = 0.005) and switched memory (p = 0.016 and p = 0.031) was detected. But transitional B lymphocytes, on the contrary, were increased in relative quantity (p = 0.003). There were no differences between the groups in the level of plasmablasts, activated B-lymphocytes CD21lowCD38low, IgM only B-cells (p > 0.05). The ROC analysis showed that the most diagnostically and prognostically significant indicators are the relative number of CD45RA+ naive T cells (cut-off ≤ 28.7%), switched memory B cells – relative (cut-off ≤ 5.0%) and the absolute number (cut-off ≤ 11 cells/µL) and the relative number of transitional B cells (cut-off ≥ 12.9%). Our data confirm the important role of regular immunophenotyping and especially subpopulations of CD45RA+ naive T cells, switched memory B cells and transitional B cells in predicting autoimmune complications in this category of patients.

Allergic diseases of the respiratory tract (allergic rhinitis and bronchial asthma) are the leading pathology in therapeutic practice. The results of a meta-analysis in the East Asian region showed that the overall prevalence of bronchial asthma and allergic rhinitis with bronchial asthma was 10.17% and 38.97%, respectively. On the territory of Mongolia there are 2883 plant species belonging to 662 genera and 128 families. In this regard, given the climatic and geographical features of Mongolia, the flowering and pollination season is long, which affects the quality of life of patients with allergic diseases. The purpose of the study is to analyze the prevalence and incidence of allergic rhinitis and bronchial asthma in Mongolia and present the experience of treatment with Antipollin.The study was conducted in Ulaanbaatar, Darkhan and Sukhbaatar for 2019-2022. Based on the data obtained, the following statistical indicators were calculated: incidence (cumulative incidence) per 1000 patients, prevalence (prevalence) for specific annual indicators and age groups.The cumulative incidence of allergic rhinitis for the fourth quarter of 2022 was determined to be from 28.9‰ to 43.92‰, the cumulative incidence of bronchial asthma was from 20.16‰ to 37.2‰, while in some age subgroups this figure exceeds 50‰. Determined that for 2022 in Ulaanbaatar – 4.39% and 2.66%, in Darkhan – 2.89% and 2.02%, in Sukhbaatar – 2.91% and 3.72% of patients were diagnosed with allergic rhinitis and bronchial asthma, respectively. In the cities of Ulaanbaatar and Darkhan, the number of patients registered with allergic rhinitis is 1.5 times higher than the number of patients with bronchial asthma, but in the city of Sukhbaatar the opposite trend is observed. The prevalence of bronchial asthma in the city of Sukhbaatar is much higher than in other cities, higher than in the AR, and as the population ages, this indicator only increases in the group 0 – 10 years – the prevalence of bronchial asthma is 12.72‰, which 2 times higher than the figure in Darkhan – 6.9‰ and 1.5 times higher than in Ulaanbaatar – 8.1‰.Taking into account the climatic, geographic, population and epidemiological characteristics of Mongolia, the use of Antipollin for sublingual immunotherapy is the optimal solution that can improve the quality of life of thousands of patients suffering from allergic diseases of the respiratory tract.

TLR2 is an exceptional pattern-recognizing receptor because of its ability to heterodimerise with different types of TLRs, which allows it to recognize a wide range of molecular structures on the surface of pathogens. Polymorphisms in genes involved in the TLRs signaling cascade may be a factor in host susceptibility to the development of inflammation, affecting the outcome of a number of infectious diseases and immune diseases. The variant Arg753Gln (rs5743708) in the TLR2 gene is the most characterized missense mutation of the coding region in the TIR domain, which involves the substitution of arginine for glutamine at position 753 of the protein sequence. This functionally significant substitution leads to altered signaling and is associated with inflammatory responses. In this study, we investigated the association of the Arg753Gln (rs5743708) polymorphism of the TLR2 gene with the level of its expression in nonagenarians. The study included 82 nonagenarians. Frailty was detected in 41 subjects using a short physical performance battery, with registration in the test ≤ 7 points. It was shown that carriage of the Gln allele is statistically significantly associated with an increased risk of developing frailty; patients with the Arg/Gln genotype have a 12.8-fold higher chance of developing this geriatric syndrome. The Arg allele and the Arg/Arg genotype were found to be protective factors in the development of frailty in nonagenarians. Analysis of TLR2 gene expression in nonagenarians revealed a 2.79-fold increase in TLR2 expression relative to donors. Evaluation of TLR2 gene expression level in groups of nonagenarians with the presence and absence of frailty showed a 1.4-fold increase in TLR2 gene expression in nonagenarians with this geriatric syndrome. In patients with the Arg/Gln genotype, TLR2 gene expression was 1.3 times higher than in the group with the Arg/Arg genotype and 1.6 times higher than in the group with the Gln/Gln genotype. The increased frequency of occurrence of the Arg/Gln genotype of the Arg753Gln polymorphism of the TLR2 gene in nonagenarians with frailty may be due to increased gene expression of this receptor. It is necessary to conduct further functional and molecular genetic studies.

CD3+CD20+T lymphocytes are a population of T cells that, along with standard T cell markers, express the atypical membrane molecule CD20 (a traditional B cell marker). These cells were identified not so long ago and are currently being actively studied. Normally, they constitute up to 3-5% of the CD3+T cell compartment in human peripheral blood, and are also found in primary and secondary lymphoid organs, cerebrospinal fluid, brain tissue and liver. In healthy individuals, CD3+CD20+T cells are heterogeneous and contain a lower proportion of CD4+ cells, but produce higher levels of GM-CSF, IFNγ, IL-17, TNFα, IL-4, IL-10, adhesion molecules and chemokine receptors than CD3+CD20-T cells, indicating a highly activated proinflammatory phenotype with properties potentially promoting their pathogenic infiltration into the CNS. Recent studies have established the pathogenic behavior of CD3+CD20+T cells in a wide range of diseases, including hematological and non-hematological CD20+T cell malignancies and HIV, as well as autoimmune pathologies, in particular multiple sclerosis, a disabling inflammatory neurodegenerative disease that is accompanied by damage to the myelin sheath nerve fibers. CD20 positive T cells are detected in patients with multiple sclerosis in the peripheral blood, cerebrospinal fluid (occur at a frequency similar to that of B cells and show a correlation with disease severity) and white matter of the brain. CD20 positive T lymphocytes in the peripheral blood of patients with multiple sclerosis have been shown to produce high levels of IFNγ and IL-17А, which are two proinflammatory cytokines involved in the pathogenesis of this disease. It is possible that CD20+T cells represent a separate subpopulation of Th17 cells, the so-called Th1-polarized Th17, which are the product of redifferentiation of Th17 cells into Th1 and combine the phenotypic characteristics of both populations. And the expression of CD20 T cells may be a valuable marker that determines the target subpopulation of such pathogenic T cells, as well as serve as a target for therapy of autoimmune diseases.

Heat shock proteins of the 70 kDa family (HSP70) are intracellular chaperones necessary for the cell to maintain protein homeostasis. In the cytosol, under normal conditions, these proteins promote the correct folding of proteins, preventing their aggregation, and are involved in protein transport and cell survival. Among the HSP70, there is a pool of stress-inducible proteins Hsp70, which significantly increases in response to a number of stress factors and facilitates cell recovery after stress. Tumor cells, unlike normal, are characterized by the ability to present Hsp70 on the surface of the cell membrane. Membrane-bound Hsp70 can be considered as a danger signal and enhance or inhibit immune responses. A three-dimensional model of cells in the spheroids in varying degrees simulates the structural organization of solid tumors. In cultures of multicellular spheroids (3D), hypoxia and nutrient gradients are formed within the spheroids, which can affect the translocation of Hsp70 to the cell membrane. The purpose of this work was a comparative analysis of Hsp70 expression on tumor cells of various origins when cultivated in a monolayer state (2D) and 3D cultures. Analysis was carried out on breast and pancreatic tumor cell lines, colon and prostate carcinomas, and lymphomas using flow cytometry and confocal microscopy methods. Cultivation in 3D cultures was performed using the antiadhesive PolyHEMA substrate. The results showed that not all carcinomas from our panel express Hsp70 in both 2D and 3D cultures. Some tumor lines have membrane Hsp70 only in 3D cultures. Hsp70 expression was detected on: BT20 breast cancer cells; colon carcinoma SW837; pancreas PANC1; and prostate PC-3. Analysis of Hsp70-positive carcinomas of various localizations in 2D and 3D models may be useful for the application of antibodies against Hsp70 as a vector for the delivery of anticancer drugs.

The transcription factor STAT3 serves as an immunoregulator by playing a crucial role in cytokine receptor signaling. However, in various cancer cell types, STAT3 is involved in the molecular mechanisms of oncogenesis. Specifically, in glioblastoma, the STAT3 immunoregulator has been linked to resistance to temozolomide, the most commonly used chemical agent for treating this type of cancer. Furthermore, literature suggests that activation of this oncogene in glioblastoma cells can significantly impact the tolerogenic tumor microenvironment, weakening the antitumor immune response and contributing to the aggressive course of the disease. Therefore, suppressing STAT3 may not only affect cell growth and resistance to chemotherapy but also enhance the immune response by improving the tumor microenvironment.The development of sequencing technologies has revealed that most of the transcribed material in the cell is noncoding. Long non-coding RNAs are gaining popularity in the study of oncogenesis due to their functional role in the development of various diseases, including oncology. A subtype of long non-coding RNAs transcribed from enhancer elements, known as enhancer RNAs, has garnered attention due to their high specificity in various cells and tissues. Gene co-expression analysis in glioblastoma tumors showed a correlation between STAT3 expression and the enhancer RNA LINC00910, which is located in the same chromosomal domain as the Stat3 gene. Previous literature has shown that LINC00910 is associated with both colorectal and gastric cancer. Additionally, data from the GeneHancer database suggests that the enhancer RNA LINC00910 may be involved in regulating the STAT3 immunoregulator. RNA interference was used to effectively knockdown the enhancer RNA LINC00910, resulting in an 8- to 10-fold reduction in its expression in glioblastoma cell lines. The reduction of LINC00910 expression did not significantly affect Stat3 gene expression in glioblastoma cell lines DBTRG-05MG and U251. This suggests that the correlation between LINC00910 RNA expression and STAT3 gene expression is not due to LINC00910’s direct involvement in STAT3 gene regulation in these cells. Further studies using the selected interfering RNA will help to clarify the role of the enhancer RNA LINC00910 in other signallingsignaling pathways, as well as its potential relationship with cancer development.

One of the urgent problems of medicine is to clarify the pathogenetic mechanisms of glomerulonephritis (GN) with refractory nephrotic syndrome (NS). In 30% of cases, refractory NS has a genetic nature. The role of human histocompatibility system (HLA) genes in the development of refractory NS has not been sufficiently studied. The purpose of this study was to study the association of two-locus haplotypes of HLA class II gene alleles with GN manifested by refractory NS. The typing of HLA class II genes in 136 patients with NS was performed by polymerase chain reaction (PCR), which included the identification of 13 alleles of the DRB1, 8 – DQA1, and 12 – DQB1 loci. The cohort of the examined patients was divided into two groups: a group of patients with refractory NS and a group of patients with rare relapses, with a lack of refractoriness to the therapy. Persons of Chuvash nationality were selected for the study. In the studied groups of patients, the values of the nonequilibrium coupling of alleles (D) were determined to identify characteristic two-locus haplotypes and their frequency according to the formulas of Piazza A. and coauthors. To assess the association of refractory NS with HLA haplotypes, relative risk values (RR) were calculated using the formula Woolf B. and Haldane J. The statistical significance of the association was assessed using the twosided Fisher exact method for four-field tables (PF). The highest value of RR was found in the haplotype HLA-DRB1*11(05)-DQA1*0301. Its value was 42.1 (PF = 0.005). Another statistically significant value was the RR value of the haplotype HLA-DRB1*15(02)-DQB1*0602-8, equal to 0.2 (PF = 0.004). As a result of the study, the haplotype DRB1*11(05)-DQA1*0301, associated with an increased risk of refractory NS, and the protective haplotype DRB1*15(02)-DQB1*0602-8, reducing the risk of refractory NS were found in the HLA genotype of individuals in the Chuvash population.

Next generation sequencing (NGS) allows high-resolution and allelic HLA-typing. The most common platform is MiSeq (Illumina, USA). Currently, the maintenance of this device is difficult, which has necessitated the search for analogues that are not inferior in capacity and quality of HLA-typing. The purpose of our study was a comparative analysis of the results of HLA-typing by NGS using the MiSeq (Illumina, USA) and FastaSeq 300 (Gene-Mind, China) platforms. The study included DNA samples of hematopoietic stem cell (HSC) donors: 12 samples were obtained and analyzed at the FSBI RosNIIGT FMBA of Russia and 24 samples at the National Medical Research Center for Hematology of the Ministry of Health of the Russian Federation. HLA typing of all samples was performed twice: using a MiSeq sequencer and a FastaSeq 300 sequencer. The results of HLA-typing of 12 samples from the FSBI RosNIIGT FMBA of Russia, obtained on two platforms, coincided. DRB1 allele of one sample was achieved as group P with MiSeq, but as group G with FastaSeq 300. The results of HLA-typing of 24 samples from the National Medical Research Center for Hematology of the Ministry of Health of the Russian Federation, obtained on two platforms, coincided. However, differences in the level of resolution of HLA-typing were observed for 10 samples. A higher level of resolution with using MiSeq was observed for genes: B – 2 cases; C, DRB3, DRB4 – 3 cases each. When using FastaSeq 300, a higher level of resolution was achieved a little more often and was established for the genes: DRB1 – 4, DQB1 – 7, DPA1 – 10 cases. Differences in the level of resolution of HLA-typing for some samples are not critical, since high-resolution typing results are currently used to select HSC donor-recipient pairs. Our study indicated the possibility of effectively using the FastaSeq 300 for HLA-typing.

Type I allergy is mediated by the formation of IgE antibodies to proteins secreted by nonreplicating microorganisms (plant pollen, house dust mites, etc.) that enter the mucous membranes in very low concentrations. The mechanisms and localization of naive B cells’ switching to IgE production have not been fully determined. The aim of this work was to determine the switching site of B cells and the traffic of IgEproducing B cells in mice immunized with a low dose of equimolar mixture of egg proteins Gal d1, Gal d2, and Gal d3. Allergens in saline solution were injected into the withers of mice 9-10 times with an interval of 2-3 days; the total dose was 2.7 µg/mouse. The production of IgE to Gal d proteins in the blood and by B cells isolated from the withers, draining lymph nodes, spleen, and bone marrow of immune mice was analyzed in dynamics after cessation of sensitization. Both in blood and in in vitro cultures, the dominance of IgE changed from the recognition of the LMW Gal d2 during the sensitization of mice to the HMW Gal d3 after sensitization was discontinued. In this model, an IgG memory response appeared only a month after the end of sensitization and recognized only Gal d3. In vitro cultures showed that B cells switched to IgE production locally in the withers with low traffic to the spleen. In the blood serum, IgE titers for all Gal d proteins decreased after the cessation of sensitization and persisted for a long time. A month after the cancellation of the sensitization, a pool of B cells producing IgE in vitro appeared in the spleen. These B-cells died after 20-30 days as no in vitro IgE production was observed later than 85-90 days. The results obtained allowed us to draw several conclusions. B cells switch to IgE synthesis locally at the site of allergen injections. The response was two-phase: LMW Gal d2 was recognized in the early response, while HMW Gal d3 was recognized in the late phase. In this model, the IgG response to HMW Gal d3 was clearly dominant. In conclusion, it has been shown that when the immune system recognizes a mixture of proteins originating from some allergen, the dominance of proteins recognized by both IgE and IgG is observed. Since allergy patients most often do not have IgG antibodies, it can be assumed that in this case an acute phase response, supported by antigen intake, is observed, in which LMW allergens are recognized.

Despite the success in early diagnosis and drug treatment of breast cancer patients, long-term treatment results are still unsatisfactory, and in this regard, the role of research aimed at studying various mechanisms of development of this disease is increasing. It has been established that the degree of tumor infiltration by immune cells and their composition are directly related to the development of the disease and the response to therapy. Multiparameter flow cytometry (PC) allows studies of the subpopulation composition of TILs. The aim of the work was to study the features of the lymphoid microenvironment (TILs) of tumors in patients with primary operable and locally advanced breast cancer by PC. The study included patients with primary operable breast cancer (group 1, n = 121) and locally advanced (group 2, n = 80) receiving treatment at the N. N. Blokhin NMRCO. The TILs of a tumor obtained intraoperatively or by a core-biopsy were examined. The patients were divided into 3 subgroups: 1 subgroup – the degree of infiltration up to 1%, 2 subgroup – the degree of infiltration from 1 to 10%, and 3 subgroup – the degree of infiltration over 10%. Patients of group 1 were characterized by high functional activity and concentration of effector cells with a low degree of tumor infiltration, and as the number of lymphocytes in the tumor increased, an increase in the pool of CD4+ cells and CD4 Treg was noted simultaneously with a decrease in the number and functional activity of effector TILs. In group 2 were no significant differences in the cellular composition of TILs in subgroups with a weak and moderate degree of infiltration, and a variant with infiltration of more than 10% was recorded in only one patient. The lack of association of the TILs subpopulation structure in subgroups with varying degrees of tumor infiltration indicates a similar nature of the local immune response in locally advanced breast cancer.Thus, unlike patients with locally advanced breast cancer, patients with primary operable breast cancer show a change in the type of local immune response from effector to regulatory. A high degree of tumor infiltration is characterized by depletion of effector cell function.

All types of immune cells are involved in the pathogenesis of multiple myeloma (MM). Granulocytic (G-MDSCs) and monocytic myeloid-derived suppressor cells (M-MDSCs) have significant protumor effects. The T cell immune response may be reduced due to the development of T cell exhaustion, characterized by the expression of inhibitory receptors PD-1, TIM-3, etc. Granulocyte colony-stimulating factor (G-CSF) supports the generation and expansion of MDSCs and can influence the functional properties of T cells. The purpose of our work was to investigate the possible effect of stimulation with G-CSF drugs on the induction of PD-1 and TIM-3 expression by T cells in patients with MM. The study included 40 patients with MM who underwent mobilization of hematopoietic progenitor cells with G-CSF drugs (5 mcg/kg/day) for 4-5 days. Content of CD4+PD-1+, CD4+TIM-3+, CD8+PD-1+, CD8+TIM-3+T cells, Lin-HLA-DR-CD33+CD66b+G-MDSCs, and CD14+HLA-DR-M-MDSCs was assessed before the start of a course of G-CSF injections (n = 33), after a course of G-CSF on the first day of separation of hematopoietic progenitor cells (n = 28) and after 3-6 months (n = 40) by flow cytometry. The relative content of G-MDSCs and M-MDSCs was significantly higher in patients with MM after a course of G-CSF. After 3-6 months, the content of G-MDSCs and M-MDSCs decreased to the initial values. After the course of G-CSF, an increase in the content of CD4+PD-1+T cells was noted compared to the values before the study. After 3-6 months, the content of this population did not differ from the initial values. The relative numbers of CD4+TIM-3+, CD8+PD-1+, and CD8+TIM-3+T cells did not change after a course of G-CSF. There were no significant correlations between the content of the populations of MDSCs and T cells expressing PD-1 and TIM-3 after a course of G-CSF.Mobilization of hematopoietic stem cells by G-CSF in patients with MM is accompanied by a transient increase in MM populations and an isolated increase in CD4+PD-1+T cells.

Innate immune cells are important participants in inflammatory and fibrotic processes in systemic scleroderma (SSc). The pathogenesis of SSc involves immune cells, primarily macrophages, whose disorders are based on mitochondrial cell dysfunction. Mitochondrial DNA (mtDNA) copy number is used as a surrogate marker of mitochondrial cell dysfunction. The aim of the study was to evaluate the number of mtDNA copies in CD14+ monocytes and in all cell populations circulating in the blood in patients with SSc compared to healthy controls.The study included 25 patients with SSc (22 women and 3 men, median age 49 (43-57) years and disease duration 4.6 (1.0-9.6) years) and 25 people without autoimmune diseases or chronic inflammatory diseases matched by age and gender. The majority of patients (80%) had a limited form of SSc. All study participants did not receive antirheumatic therapy. DNA was isolated from CD14+ monocytes and whole blood. Absolute mtDNA copy number was measured using digital PCR. The number of mtDNA copies per cell used for analysis was calculated as the ratio of mtDNA and nDNA copies.It was found that in patients with SSc, the number of mtDNA copies in CD14+ monocytes was higher (108 (60-162) vs 72 (59-79), p = 0.01), and the indicator of all cell populations circulating in the blood did not differ in compared with the control group (109 (72-171) and 128 (85-227), p = 0.17). A negative relationship was found between the number of mtDNA copies and the duration of the disease, and a positive relationship with LPS-stimulated IL-6 secretion by cultured CD14+ monocytes.The study results suggest that increase of mtDNA copy number in CD14+ monocytes is a possible mechanism to maintain the reduced function of defective mitochondria in monocytes from patients with SSc associated with the development and progression of SSc.

Macrophages are key regulatory cells of fibrogenesis. They can have pro- or antifibrotic activity due to their plasticity and heterogeneity. Some studies have shown the antifibrotic effect of macrophages on dermal fibroblasts, but the effect of macrophages on the lung fibroblast functions remains unexplored. Therefore, the purpose of this study was to examine the influence of conditioned media of human macrophage differentiated by M-CSF/GM-CSF and further dexamethasone polarized/unpolarized on the TGF-β-induced lung fibroblast differentiation. Macrophages were derived from peripheral blood monocytes of healthy donors. Monocytes were differentiated by M-CSF or GM-CSF for 7 days. On day 5, dexamethasone was added to generation of polarized macrophages M-M(Dex) and GM-M(Dex). Polarized macrophages were compared with non-polarized M-M0 and GM-M0, to which dexamethasone was not added. Next, the conditioned medium of these macrophage subtypes was collected and tested for inhibition the lung fibroblast differentiation (HLF210 cell line). To do this, TGF-β (inducing differentiation factor) and conditioned macrophage medium were added to fibroblast cultures. Effectiveness of differentiation was estimated by the expression of the myofibroblast marker, α-smooth muscle actin (α-SMA), and the production of extracellular matrix protein, collagen I. The expression of α-SMA was determined using flow cytometry. The concentration of collagen I was measured by ELISA. Since our data indicates that spontaneous activation of fibroblasts occurs during standard cultivation, the α-SMA expression was investigated in 3D culture of fibroblasts. Notably, the content of α-SMA-positive cells in 3D cultures was significantly reduced, indicating more physiological growth cells. Regardless of the differentiation stimulus, the conditioned media of dexamethasone-polarized macrophages do not affect the level of collagen I production or the α-SMA expression. On the contrary, M-M0 showed a strong inhibitory effect that reduced the amount of collagen I in the fibroblast cultures and the expression of marker myofibroblasts by fibroblasts. Interesting, GM-M0 had no such effect and did not prevent lung fibroblast differentiation like polarized cells. Taken together, the findings suggest that M-M0 macrophages may have antifibrotic properties. Furthermore, the lack of this effect in GM-M0 macrophages indicates that the differentiation factor plays a significant role in the development of the antifibrotic macrophage phenotype.

Malignant transformation of lymphopoiesis in lymph nodes (LN) is accompanied by structural rearrangement of the LN stroma and changes in the glycosylation of membrane and cytoplasmic proteins. For the histochemical detection of transforming lymphoid cells and remodeled LN stroma, we used the tomato lectin Lycopersicon esculentum, which is able to bind to surface and cytoplasmic glycoproteins of the majority of LN cells. The study aimed to investigate the characteristics of cell architectonics with a high level of protein glycosylation in the LN of patients with B-chronic lymphocytic leukаemia (B-CLL). The study material were biopsy specimens of supraclavicular and cervical LNs from patients of the First Republican Clinical Hospital of the Ministry of Health of the Udmurt Republic with a confirmed diagnosis of B-CLL (16 patients), aged 49-73 years, obtained prior to treatment with their informed voluntary consent. LN biopsies from the same body regions of 12 individuals aged 48-70 years with reactive LN hyperplasia served as control samples. Paraffin sections of 7 µm thick LN were stained with FITC-conjugated tomato lectin and fluorescent dye propidium iodide (IP) and examined using a Nikon Eclipse200 microscope equipped with a luminescence unit and digital camera. Analysis of LN preparations from patients with B-CLL revealed significant changes in the histotopography of cells and extracellular structures with a high degree of glycosylation. Follicles in the cortex were replaced by an array of small lymphocytes against a background of proliferating centers containing lymphocytes with dispersed packing of IP-labelled chromatin. In this area we also observed a uniform network of thin lectin-labelled reticular fibres and a large number of small blood vessels. Macrophage-like cells, clearly identifiable in the germinal centres of follicles in control, were absent in B-CLL. Their increased number and intensity of luminescence was observed in the subcapsular sinus area and in the paracortical area around collagen bundles formed by conduits, as well as around connective tissue trabeculae of the brain substance. The differences observed in the histological topography of highly glycosylated LN cells in B-CLL suggest that the proposed staining method is informative and facilitates the diagnosis of this disease in histological studies.

Tumor-associated macrophages (TAMs) are an important and most represented population of immune cells in the tumor microenvironment. To a great extent, TAMs can determine the direction of the antitumor immune response; they can either additionally stimulate it or on the contrary contribute to the formation of immunosuppressive microenvironment. At the same time, under the influence of tumor cells and antitumor therapy, many cells in the tumor microenvironment (TME) can develop a state of senescence. Over the last decade, the topic of senescence and the search for therapies aimed at removing senescent cells has gained popularity. In the search for new therapeutic strategies to treat cancer, senescent cells of the immune system in the tumor microenvironment have received special attention since the presence of senescent TAMs in tumors is associated with poor prognosis and poor response to therapy. Given the relevance of studying the role of senescent immune cells in TME (in particular tumor-associated macrophages), we performed a comparative analysis of experimental protocols to obtain tumor-associated macrophages in vitro to determine the most relevant approach. We tested two protocols for obtaining macrophages from mouse bone marrow: (1) by adding conditioned medium from the L929 mouse sarcoma cell line (LCCM) (LCCM-BMDM); and (2) by adding recombinant mouse M-CSF (M-CSF-BMDM). We showed that LCCM-BMDMs, compared to M-CSFBMDMs, have increased expression of the arginase enzyme (Arg1), which can inhibit the activity of anti-tumor cytotoxic lymphocytes by depleting arginine in the tumor microenvironment. LCCM-BMDMs also exhibited increased secretion of factors characteristic of the senescence-associated secretory phenotype (SASP): IL-6 and TNF. Both Arg1 and IL-6 and TNF are markers characteristic of senescence-associated macrophages. Thus, the use of LCCM to obtain primary macrophage culture limits further steps in creating a model of tumor-associated macrophages that reflects the specific characteristics of the macrophage phenotypic response for different tumor types aAnd also limits studies of senescence formation in tumor-associated macrophages in models of carcinogenesis other than sarcoma. We believe that differentiation of macrophages in the presence of M-CSF appears to be a more preferable protocol to study TAMs and senescent TAMs to test new therapeutic strategies.

The paper presents a description of a clinical case of Gougerot–Hailey–Hailey disease with proven dysregulation of the immune response. The disease, also known as chronic benign familial pemphigus, is a rare genetic disorder that causes bullous skin lesions with intraepidermal localization. It is named after the authors who first described it. In most cases, Gougerot–Hailey–Hailey disease is caused by a mutation in the ATP2C1 gene that disrupts calcium regulation. Calcium accumulation alters normal intercellular adhesion in the skin, resulting in characteristic bullous skin lesions. Gougerot–Hailey–Hailey disease is inherited in an autosomal dominant pattern. The frequency of distribution in the population is 1:50000 people. However, the variety of clinical phenotypes of the disease without a proven hereditary predisposition and the absence of a mutation in this gene allows us to assert the presence of other pathogenic factors, in particular, disorders in the immune system. In the clinical case we are describing, no clear genetic predisposition was revealed, but serious disorders in the immune system were detected. In particular, TNK cells are virtually non-existent. Analysis of subpopulations of natural killer cells indicates a decrease in the relative and absolute numbers of NK cells expressing CD16 and CD56 antigens, NK cells with high cytolytic activity, NK cells expressing the α chain of the CD8 antigen and having the ability to repeatedly perform their cytolytic function. When assessing B cell subpopulations, there is an increase in the relative content of B2 cells and B1 cells associated with production of autoantibodies, in parallel with an increase in the percentage of regulatory T helper cells with immunosuppressive function. Based on the identified changes in the immune system, we performed immunomodulatory therapy: discrete plasmapheresis, 5 procedures in 2 days, then intravenous drip injection of normal human immunoglobulin 25 mL (50 μg/mL) in 3 days, 5 infusions. The dynamics of cutaneous manifestations indicates a moderate positive effect. Obviously, further in-depth study of immune parameters in Gugerout-Haley-Haley disease, presumably in the innate immune system, is required. Probably, success in treatment and achievement of stable remission will be possible with the rational selection of anticytokine therapy.

It is known that chronic social stress leads to immunity disorders in humans and experimental animals. It has been shown that the effect of stress is also manifested in changes in the level of expression of genes involved in the functioning of various physiological systems in the brain of mice, in particular, in the hypothalamus. It was noted that in stressed animals, genes involved in the processes of carcinogenesis and apoptosis change their expression, and in animals without signs of developing a malignant process, but under conditions conducive to tumor growth. In this regard, we used the RNA-seq method to study the expression of cytokine response genes in the hypothalamus of male mice under the influence of chronic social stress caused by repeated experience of defeats in intermale confrontations, compared with control individuals. Multidirectional changes in the expression of cytokine genes, their receptors and genes performing a regulatory function were detected (IL17d, IL18, IL33, Csf1r, Csf2ra, IL11ra1, IL13ra1, IL2ra, IL3ra, IL5ra, Lifr, Cish, IL4i1, Irf1, Irf5, Irf9, Jak2, Socs3, Stat3, Tgfb1, Tlr3). Thus, it has been shown that changes in the cytokine response in the brain under the influence of stress occur at the level of changes in gene expression. In this case, we should not talk about the activation of the system or a decrease in its activity, but about the disruption of its functioning. Next, we analyzed the correlations between the level of expression of genes of the cytokine system and the main genes of carcinogenesis and apoptosis that we studied earlier (Akt1, Bag6, Foxp4, Mapk3, Mapk8, Nol3, Pdcd10, Xiap). The Akt1, Jak2, Stat3 genes were identified, for which the maximum number of correlations was found, moreover, negative correlations were most characteristic of Jak2, and positive correlations were most characteristic of Stat3 and Akt1. In addition, protein-protein interactions between genes of carcinogenesis and apoptosis and genes of the cytokine system were analyzed using the String database in mice under chronic social stress. It was confirmed the key role of these genes in the development of dysfunction of cytokines in the brain.

Abstract Although Omicron RBD of SARS-CoV-2 accumulates many mutations, the backbone region (truncated RBD) of spike protein is highly conserved. Here, we designed several subunit vaccines by keeping the conserved spike backbone region of SARS-CoV-2 Omicron BA.1 subvariant (S-6P-no-RBD), or inserting the RBD of Delta variant (S-6P-Delta-RBD), Omicron (BA.5) variant (S-6P-BA5-RBD), or ancestral SARS-CoV-2 (S-6P-WT-RBD) to the above backbone construct, and evaluated their ability to induce immune responses and cross-protective efficacy against various SARS-CoV-2 variants and SARS-CoV. Among the four subunit vaccines, S-6P-Delta-RBD protein elicited broad and potent neutralizing antibodies against all SARS-CoV-2 variants tested, including Alpha, Beta, Gamma, and Delta variants, the BA.1, BA.2, BA.2.75, BA.4.6, and BA.5 Omicron subvariants, and the ancestral strain of SARS-CoV-2. This vaccine prevented infection and replication of SARS-CoV-2 Omicron, and completely protected immunized mice against lethal challenge with the SARS-CoV-2 Delta variant and SARS-CoV. Sera from S-6P-Delta-RBD-immunized mice protected naive mice against challenge with the Delta variant, with significantly reduced viral titers and without pathological effects. Protection correlated positively with the serum neutralizing antibody titer. Overall, the designed vaccine has potential for development as a universal COVID-19 vaccine and/or a pan-sarbecovirus subunit vaccine that will prevent current and future outbreaks caused by SARS-CoV-2 variants and SARS-related CoVs.

Abstract The development of broad-spectrum coronavirus vaccines is essential to prepare for future respiratory virus pandemics. We demonstrated broad neutralization by a trivalent subunit vaccine, formulating the receptor-binding domains of SARS-CoV, MERS-CoV, and SARS-CoV-2 XBB.1.5 with Alum and CpG55.2. Vaccinated mice produced cross-neutralizing antibodies against all three human Betacoronaviruses and others currently exclusive to bats, indicating the epitope preservation of the individual antigens during co-formulation and the potential for epitope broadening.

Abstract Objectives The pathogenic microorganisms that cause intestinal diseases can significantly jeopardize people's health. Currently, there are no authorized treatments or vaccinations available to combat the germs responsible for intestinal disease. Methods Using immunoinformatics, we developed a potent multi-epitope Combination (combo) vaccine versus Salmonella and enterohemorrhagic E. coli. The B and T cell epitopes were identified by performing a conservancy assessment, population coverage analysis, physicochemical attributes assessment, and secondary and tertiary structure assessment of the chosen antigenic polypeptide. The selection process for vaccine development included using several bioinformatics tools and approaches to finally choose two linear B-cell epitopes, five CTL epitopes, and two HTL epitopes. Results The vaccine had strong immunogenicity, cytokine production, immunological properties, non-toxicity, non-allergenicity, stability, and potential efficacy against infections. Disulfide bonding, codon modification, and computational cloning were also used to enhance the stability and efficacy of expression in the host E. coli. The vaccine's structure has a strong affinity for the TLR4 ligand and is very durable, as shown by molecular docking and molecular modeling. The results of the immunological simulation demonstrated that both B and T cells had a heightened response to the vaccination component. Conclusions The comprehensive in silico analysis reveals that the proposed vaccine will likely elicit a robust immune response against pathogenic bacteria that cause intestinal diseases. Therefore, it is a promising option for further experimental testing.

Abstract Background TLR7 is a key player in the antiviral immunity. TLR7 signaling activates antigen-presenting cells including DCs and macrophages. This activation results in the adaptive immunity including T cells and B cells. Therefore, TLR7 is an important molecule of the immune system. Based on these observations, TLR7 agonists considered to become a therapy weaponize the immune system against cancer. Radiation therapy (RT) is one of the standard cancer therapies and is reported to modulate the tumor immune response. In this study, we aimed to investigate the anti-tumor activity in combination of TLR7 agonist, DSP-0509, with RT and underlying mechanism. Result We showed that anti-tumor activity is enhanced by combining RT with the TLR7 agonist DSP-0509 in the CT26, LM8, and 4T1 inoculated mice models. We found that once- weekly (q1w) dosing of DSP-0509 rather than biweekly (q2w) dosing is needed to achieve superior anti-tumor activities in CT26 model. Spleen cells from the mice in RT/DSP-0509 combination treatment group showed increased tumor lytic activity, inversely correlated with tumor volume, as measured by the chromium-release cytotoxicity assay. We also found the level of cytotoxic T lymphocytes (CTLs) increased in the spleens of completely cured mice. When the mice completely cured by combination therapy were re-challenged with CT26 cells, all mice rejected CT26 cells but accepted Renca cells. This rejection was not observed with CD8 depletion. Furthermore, levels of splenic effector memory CD8 T cells were increased in the combination therapy group. To explore the factors responsible for complete cure by combination therapy, we analyzed peripheral blood leukocytes (PBLs) mRNA from completely cured mice. We found that Havcr2low, Cd274low, Cd80high, and Il6low were a predictive signature for the complete response to combination therapy. An analysis of tumor-derived mRNA showed that combination of RT and DSP-0509 strongly increased the expression of anti-tumor effector molecules including Gzmb and Il12. Conclusion These data suggest that TLR7 agonist, DSP-0509, can be a promising concomitant when used in combination with RT by upregulating CTLs activity and gene expression of effector molecules. This combination can be an expecting new radio-immunotherapeutic strategy in clinical trials.

Abstract Background The systemic inflammatory syndrome called “cytokine storm” has been described in COVID-19 pathogenesis, contributing to disease severity. The analysis of cytokine and chemokine levels in the blood of 21 SARS-CoV-2 positive patients throughout the phases of the pandemic has been studied to understand immune response dysregulation and identify potential disease biomarkers for new treatments. The present work reports the cytokine and chemokine levels in sera from a small cohort of individuals primarily infected with SARS-CoV-2 during the first wave of the COVID-19 pandemic in Milan (Italy). Results Among the 27 cytokines and chemokines investigated, a significant higher expression of Interleukin-9 (IL-9), IP-10 (CXCL10), MCP-1 (CCL2) and RANTES (CCL-5) in infected patients compared to uninfected subjects was observed. When the change in cytokine/chemokine levels was monitored over time, from the hospitalization day to discharge, only IL-6 and IP-10 showed a significant decrease. Consistent with these findings, a significant negative correlation was observed between IP-10 and anti-Spike IgG antibodies in infected individuals. In contrast, IL-17 was positively correlated with the production of IgG against SARS-CoV-2. Conclusions The cytokine storm and the modulation of cytokine levels by SARS-CoV-2 infection are hallmarks of COVID-19. The current global immunity profile largely stems from widespread vaccination campaigns and previous infection exposures. Consequently, the immunological features and dynamic cytokine profiles of non-vaccinated and primarily-infected subjects reported here provide novel insights into the inflammatory immune landscape in the context of SARS-CoV-2 infection, and offer valuable knowledge for addressing future viral infections and the development of novel treatments.

Abstract Background Glioblastoma is characterized by high aggressiveness, frequent recurrence, and poor prognosis. Histone acetylation-associated genes have been implicated in its occurrence and development, yet their predictive ability in glioblastoma prognosis remains unclear. Results This study constructs a histone acetylation risk model using Cox and LASSO regression analyses to evaluate glioblastoma prognosis. We assessed the model’s prognostic ability with univariate and multivariate Cox regression analyses. Additionally, immune infiltration was evaluated using ESTIMATE and TIMER algorithms, and the SubMAP algorithm was utilized to predict responses to CTLA4 inhibitor. Multiple drug databases were applied to assess drug sensitivity in high- and low-risk groups. Our results indicate that the histone acetylation risk model is independent and reliable in predicting prognosis. Conclusions Low-risk patients showed higher immune activity and longer overall survival, suggesting anti-CTLA4 immunotherapy suitability, while high-risk patients might benefit more from chemotherapy. This model could guide personalized therapy selection for glioblastoma patients.